Figure 9.

ApoE protein in conditioned media was elevated in response to complement challenge. RPE cells were primed with or without S58 (1.2 mg/mL) for 30 minutes and then treated with either 6% C1q-Dep or 6% HiC1q-Dep for 6 hours. The medium was then exchanged with serum-free medium. After media exchange, cultures were incubated for either 1, 24, 48, or 72 hours and levels of soluble ApoE in the media were determined. Total cellular lysate proteins (15 μg) and concentrated conditioned media (30 μL) were separated by SDS-PAGE. The data are representative of two separate experiments in a 62-year-old donor with ApoE phenotype E3/E3 and CFH_YY402 variant with similar results.